HT-3
RNA quality was assessed using an Agilent 2100 Bioanalyzerand tested using RNA-free agarose gel electrophoresis. After total RNA extraction, rRNA-enriched prokaryotic mRNA was removed using the Ribo-ZeroTM Magnetic Kit . Fragment buffers were then used to fragment the enriched mRNA into short fragments. The NEB Next Ultra RNA Library Prep Kit for Illumina (neb# 7530,New England Biolabs, Ipswich, MA, USA) was used to reverse-transcribe RNA into cDNA. The purified double-stranded cDNA fragment was end-repaired, polyadenylated, and Illumina sequencing primers were used. The ligation reaction was purified using AMPure XP Beads (1.0X). The ligated fragments were selected for size via agarose gel electrophoresis and polymerase chain reaction (PCR).